Delta depth to baseline [mm],
Base Formula Active Formula Benchmark
Delta circumference to baseline
[mm], (field 3)
Base Formula Active Formula Benchmark Formula
Delta negative volume to
baseline [mm3], (field 3)
Base Formula Active Formula Benchmark Formula
An exemplary claim substantiation for the firming / filling activity of a synthetic tripeptide
R. Campiche1, E. Jackson1, D. Imfeld1, E. Wandeler1, G. Laurent1, C. Wagner2, M. Roche3, P. Séroul3, M. Massironi4, S. Stroebel5, M. Gempeler1
1DSM Nutritional Products Ltd., Personal Care & Aroma, Kaiseraugst, Switzerland, 2proDERM GmbH, Schenefeld, Germany, 3Newtone Technologies, Lyon, France, 4Cutech
Biotech srl, Padova, Italy, 5InSphero AG, Schlieren, Switzerland
* correspondance to: firstname.lastname@example.org
Keywords:tripeptide, skin firming, filling, hyaluronic acid
Material & Methods
Results in vivo
Figure 5: 2.5% active containing the tripeptide
stimulated hyaluronic acid synthesis (blue
staining) in skin tissue microspheres. bFGF
served as a positive control. Shown are
representative tissue sections.
Figure 3: (A) A formulation containing 2.5% Active
led to improved firming as measured by DynaSKIN on
the cheeks of volunteers. It even outperformed a
formulation containing a market relevant anti-ageing
technology (Benchmark). *p<0.05 vs benchmark.
*p<0.05 vs baseline. Error bars represent standard
error of the mean. (B) 3D false color images of a
cheek hole generated by the DynaSKIN airstream.
Depth, but also circumference clearly improved after
29 days of active formulation. A representative
volunteer is shown. Depth and circumference result
in negative volume.
We investigated the hyaluronic acid boosting, and firming / filling activity of the synthetic tripeptide Tetradecyl
Aminobutyroylvalylaminobutyric Urea Trifluoroacetate (trade name SYN®-HYCAN) in vitro,ex vivo and in vivo. We
provide further evidence, that the tripeptide stimulates endogenous hyaluronic acid synthesis in facial skin tissue both
in vitro and ex vivo, also after UV-irradiation. In vivo we show that this leads to a filling and firming effect similar to
hyaluronic acid injections at distinct facial sites. In addition, treatment with a formulation containing the tripeptide
leads to increased cutaneous hydration, possibly due to the high water binding capacity of hyaluronic acid .
1) Heidl M, Gräub R: Nuovo tripeptide anti-età. In: Cosmetic Technology. vol. 13; 2010: 13-16.
2) Dai G et al.: Chronic Ultraviolet B Irradiation Causes Loss of Hyaluronic Acid from Mouse Dermis Because of Down-Regulation of Hyaluronic Acid
Synthases. Am J Pathol 2007, 171(5):1451-1461
3) Kaya G et al.: Hyaluronate fragments reverse skin atrophy by a CD44-dependent mechanism. PLoS Med 2006, 3(12):e493.
4) Oh JH, et al.: Intrinsic aging- and photoaging-dependent level changes of glycosaminoglycans and their correlation with water content in human skin.
J Dermatol Sci 2011, 62(3):192-201.
5) Del Bino S, et al.: Relationship between skin response to ultraviolet exposure and skin color type. Pigment Cell Res 2006, 19(6):606-614.
Tetradecyl-Dab-Val-Dab (INCI: Tetradecyl Aminobutyroylvalylaminobutyric Urea Trifluoroacetate) is a tripeptide with anti-
ageing effects in skin. In its product form it is combined with water, glycerine, and magnesium chloride (DSM product code
503833304), and marketed under the trade name SYN®-HYCAN. It thus belongs to DSM’s SYN®-peptide family. Its sequence is
derived from the activation domain of thrombospondin-1 and modeled by rational design to activate transforming growth
factor beta-1 (TGFE1). Its main activity in vitro is the stimulation of the skin’s endogenous synthesis of glycosaminoglycans,
in particular hyaluronic acid (HA), and proteoglycans, in particular decorin and lumican . There is a decrease in
glycosaminoglycans with chronological as well as photo-ageing . Dermal fillers, like collagen or hyaluronic acid fillers, are
high on the agenda for many women to counteract cutaneous atrophy  leading to facial wrinkles, loss of skin firmness, and
skin sagging. However, such fillers require beauty doctor’s visits and needle injections. This tripeptide, though, provides an
easy to use cosmetic solution. The data presented in this poster are from new clinical and pre-clinical studies. They provide
further evidence for the tripeptide’s outstanding activity for facial skin firming and moisturization  by acting as a needle-
free hyaluronic acid booster. We propose that Tetradecyl Aminobutyroylvalylaminobutyric Urea Trifluoroacetate (hereafter
called tripeptide) acts particularly in areas below the eyes, on the cheeks, and on the jawline (Fig.1).
Figure 1: (A) Face of a young woman. (B,
C) She has an augmented cheekbone area
(fields 1 and 2), firm cheeks (field 3), and
a well defined jawline (field 4). Fields 1
to 4 denote areas of filling mainly to lift
or augment the cheek area. Hyaluronic
acid filling in these four facial areas is
often done in plastic surgery to achieve a
well defined young looking contour.
In vivo study: This was a full-face, base formula controlled, randomized single center study. It took place from October 18th to
December 15th, 2017, in Schenefeld/Hamburg, Germany. We recruited female Caucasian volunteers age 41 to 60 (mean 52.9 ±4.8
years) with self-perceived sagging of the skin. We had three cohorts (Base, Active, Benchmark) consisting of 30 volunteers each.
Volunteers were instructed to apply their respective formulation twice daily to the entire face for 29 days. Instrument
measurements to assess various skin parameters were done at baseline, day 15, and day 29. Composition of the three formulations
is listed here:
Skin firmness was measured by DynaSkin (Eotech, France). An air stream is blown perpendicularly to a defined spot on the cheek.
Depth, circumference, and negative volume of the as such generated whole in the cheek are measured.
Skin filling was measured by acquisition of 3D-facial scans using the AEVA-HE device (Eotech, France). Images were processed and
using a defined algorithm positive volume was assessed.
Skin hydration was measured by Corneometer (Courage & Kazaka, Germany). Five distinct spots each on right and left cheekbone
were measured and mean values calculated.
In vitro study: We grew 3D-microtissue (microspheres) resembling human skin. They consisted of a fibroblast core (dermis) and an
outer keratinocyte layer (epidermis). The skin microspheres were treated for ten days with the active (consisting of the peptide
product) and a positive control (basicFGF). Culture medium was replaced every other day. After ten days skin microspheres were
harvested, formalin fixed and paraffin embedded. They were cut into 6micron tissue sections for staining of hyaluronic acid (by
alcian blue kit, Chroma 8GS, AppliChem GmbH, Germany).
Ex vivo study: Abdominal skin samples from a Caucasian female donor age 46 were harvested after obtaining informed consent
and following Helsinki declaration. The solar simulator adopted for daily UV-irradiation was a BIO-SUN system produced by Vilber
Lourmat (Eberhardzell, Germany). The selected UV irradiation intensity was 3 J/cm2(= 0.1 J/cm2 UVB + 2.9 J/cm2U VA )
corresponding to 40% biologically effective dose (BED) for daylight UV . Hyaluronic acid was stained by alcian blue (Alcian Blue
8GX staining kit, Sigma- Aldrich cat #A3157). Semiquantitative evaluation of staining intensity was done by Image J software
(NIH, Bethesda, US).
Untr 5 ppm 10 ppm 5 ppm 10 ppm 500 ppm
Matrixyl tripeptide SYN®-HYCAN peptide Retinoic acid
UV 3 J/cm2
Alcian blue score [%]
Hyaluronic acid content *
UVR 3 J/cm2
Baseline Day 29
10 days incubation
Hyaluronic acid content
10 ppm Tripeptide
Result ex vivo
Figure 4: A formulation
containing 2.5% Active led to
improved skin hydration as
measured by Corneometer on
the cheekbone of volunteers.
*p<0.05 vs placebo. **p<0.01 vs
baseline. Error bars represent
standard error of the mean.
Figure 6: 10 ppm tripeptide stimulated hyaluronic acid content in skin tissue ex vivo
(blue staining) after UV-irradiation. Retinoic acid served a positive control and the
tripeptide palmGHK as anti-ageing reference. *p<0.05 vs vehicle. Error bars
represent standard error of the mean.
Figure 2: (A) A formulation containing 2.5% Active led to
increased volume as measured by AEVA-HE and distance per
pixel in the area below the eyes of volunteers. A
formulation containing 2.5% Active led to a significant
increase in volume compared to base formulation. *p<0.05
(B) 3D false color images of the area below the eyes (field
1). Displayed are images of volunteers representing
approximately the mean values shown in A. (C) Volume
measured by AEVA-HE 3D-image scans. A formulation
containing 2.5% SYN-HYCAN led to a significantly increased
volume compared to baseline after 29 days. *p<0.05 vs
baseline. Error bars represent standard error of the mean.
50 ng/ml bFGF
Orange = increase in volume
Blue = decrease in volume
Result in vitro
Volunteer 36, Active Formula
Volunteer 94, Base Formula
Day 15 Day 29
Component (INCI) Active Formula Base Formula Benchmark Formula
% w/w % w/w % w/w
Base Formulation (emulsion) 97.5 100 97
Glycerin, Aqua, Butylene Glycol, Carbomer, Polysorbate 20, Palmitoyl Tripeptide-1, Palmitoyl Tetrapeptide-7 0 0 3
Glycerin, Aqua, Magnesium chloride, Tetradecyl Aminobutyroylvalylaminobutyric urea trifluoroacetate (tripeptide) 2.5 00
Base Formula Active Formula
Delta d29 to d0
Corneometer values [capacitance], (field 2)
Delta distance vs baseline
Positive volume, area below eyes (field 1)
Delta distance vs baseline [mm]
Positive volume, jawline (field 4)
2.5% Active *
Volunteer #13, Left side, (field 3)